Analysis of chromosomal integration and deletions of yeast plasmids (original) (raw)
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Department of Biochemistry, Stanford University School of Medicine
Stanford, CA 94305, USA
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Department of Biochemistry, Stanford University School of Medicine
Stanford, CA 94305, USA
Search for other works by this author on:
Department of Biochemistry, Stanford University School of Medicine
Stanford, CA 94305, USA
Search for other works by this author on:
Received:
10 January 1977
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John R. Cameron, Peter Philippsen, Ronald W. Davis, Analysis of chromosomal integration and deletions of yeast plasmids, Nucleic Acids Research, Volume 4, Issue 5, 1 June 1977, Pages 1429–1448, https://doi.org/10.1093/nar/4.5.1429
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Abstract
Plasmid DNAs from six strains of Saccharomycescerevisiae were compared. Three different plasmids were found, designated Scp 1, Scp 2 and Scp 3, with monomer lengths of 6. 19, 6.06 and 5.97 kilobases as referenced to sequenced ØX174 DNA. DNA from each of the plasmids was inserted into a lambda vector DNA. Hybrid phage containing inserted DNA of the desired size were enriched by genetic selection and their DNAs analysed by rapid techniques. All three plasmids share the same organization, two unique sequences separated by two inverted repeats, and share basically the same DNA sequences. Scp 2 and Scp 3 differ from Scp 1 by missing a unique Hpa l site and by having small overlapping deletions in the same region. The Hpa l site in Scp 1 is, therefore, in a nonessential region and suitable for insertion of foreign DNA in the potential use of the yeast plasmid as a vector. Hybridization of labelled cloned plasmid DNA to restriction fragments of linear yeast DNA separated on agarose gels showed that the plasmid DNA was not stably integrated into the yeast chromosomal DNA.
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